The goal of the proposed research is to develop a microorganism capture/biosensor system for sensitive real-time detection of airborne pathogens. A portable aerosol sampler interfaced with electrochemical microbiological sensor technology is proposed, which is capable of sample acquisition, real-time detection, and digital output. The proposed technology will serve clinical laboratories as well as an exposure indicator in high-risk environments. The specific aims of Phase I effort are: 1. Design, fabrication and demonstration of proof of concept of an integrated bioaerosol capture and real-time biosensor system, using the non-pathogenic fast-growing Mycobacterium parafortuitum as the model microorganism. 2. Demonstration of real-time detection of the vaccine substrain M. bovis bacillus Calmette-Guerin (BCG), which has many common features with the M. tuberculosis. Long-term objectives (to be proposed for Phase II effort): 1. Development of diagnostic reagents and electrochemical biosensor test protocols for real-time detection of Mycobacterium tuberculosis and other airborne pathogens: Legionella pneumophila, Streptococcus pyogenes and Aspergillus fumigatus. 2. Development of an automated airborne microbe monitoring instrument prototype with capabilities of automated sample taking, sample processing, mycobacterium detection, and digital output. 3.Development of a miniaturized airborne microbe-monitoring instrument for continuous monitoring of clinical personnel for exposure including low-level persistent exposure pathogenic mycobacteria. PROPOSED COMMERCIAL APPLICATION: Rapid diagnostics of TB infections in point-of-care facilities.